Use of adipose tissue-derived stromal stem cells in treating fistula

a technology of stromal stem cells and adipose tissue, which is applied in the direction of genetically modified cells, skeletal/connective tissue cells, prosthesis, etc., can solve the problems of incontinence, recurrence, reinfection, and incontinence, and achieves greater homogeneity and distinct phenotypes

Inactive Publication Date: 2006-03-02
AUTONOMOUS UNIVERSITY OF MADRID +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Surgery for anorectal fistulae is not without side effects, including recurrence, reinfection, and incontinence.
Management of fistulae in patients with Crohn's disease continues to present an extremely challenging problem since many such fistulae do not respond to available treatments.
Such fistulae and their recurrence are a very distressing complication that significantly reduces the quality of life of affected patients.
However, many patients are not cured.
Failure of fistulae to heal is probably due to the suboptimal quality of tissues that have been affected by Crohn's disease.
This hole results in permanent incontinence of urine and / or feces.
This calculation could be a significant underestimate.
Thus, a significant percentage of women have obstetrical fistulae that cannot be repaired surgically.

Method used

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  • Use of adipose tissue-derived stromal stem cells in treating fistula
  • Use of adipose tissue-derived stromal stem cells in treating fistula
  • Use of adipose tissue-derived stromal stem cells in treating fistula

Examples

Experimental program
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Effect test

example 1

Preparation of Stem Cells from Lipoaspirates with Improved Homozeneity

[0101] Adipose tissue was obtained by liposuction, under local anaesthesia and general sedation. A hollow blunt-tipped cannula was introduced into the subcutaneous space through a small incision (less than 0.5 cm in diameter). With gentle suction, the cannula was moved through the adipose tissue abdominal-wall compartment for mechanical disruption of the fatty tissue. A saline solution and the vasoconstrictor epinephrine were injected into the adipose tissue compartment to minimize blood loss. In this way, 80 to 100 ml of raw of lipoaspirate were obtained from each patient to be treated.

[0102] The raw lipoaspirate was washed extensively with sterile phosphate-buffered saline (PBS; Gibco BRL, Paisley, Scotland, UK) to remove blood cells, saline and local anaesthetic. The extracellular matrix was digested with a solution of type II collagenase (0.075%; Gibco BRL) in balanced salt solution (5 mg / ml; Sigma, St. Loui...

example 2

Characterization of Stem Cells from Lipoaspirates with Improved Homogeneity

[0104] To characterize the cells by immunofluorescence staining, cells were plated at low density in DMEM plus 10% FBS on glass cover slips in 24-well plates. For immunohistochemistry studies, cells were washed with PBS and fixed in acetone for 10 min at −20° C. For staining of a-actin, cells were fixed in 4% paraformaldehyde for 10 min at RT. After blocking with a PBS that contained 4% goat serum and 0.1% Triton X-100, cells were incubated at 4° C. overnight with primary antibodies against the following cell markers at the indicated dilutions [(i) alpha-actin; Dako, Glostrup, Denmark; 1 / 50; (ii) vimentin; Sigma, St. Louis, USA; 1 / 200; (iii) CD 90; CYMBUS, Biotechnology LTD, Chandlers Ford, Hants, UK; 1 / 50; (iv) Factor VIII; Dako; 1 / 100; (v) CD 34; Chemicon, Calif., USA; 1 / 100; (vi) c-Kit; Chemicon; 1 / 100; (vii) desmin; Dako; 1 / 100; (viii) cytokeratin; Dako; 1 / 100 and (ix) S-100; Dako; 1 / 50]. Then cells were...

example 3

Preparations of Stem Cells Comprising Fibrin Glue for Use in Treating Fistula

[0122] For clinical use, the cells as prepared above may be used after three or fewer passages (FIG. 3), but are preferably used after two or more passages as described above to afford a cell preparation with higher homogeneity. Cell cultures for clinical use were trypsinized for 3 min at 37° C. Trypsinization was stopped by addition of DMEM plus FBS, and the suspension was centrifuged at 110×g for 5 min. Cells were washed in PBS and the suspension was centrifuged again at 150×g for 5 min. Cell were resuspended at between 3 and 30×106 cells / ml in 1 to 2 ml of Ringer's lactate solution and put in a suitable syringe. Human serum albumin (HSA) may optionally be added to the Ringer's lactate solution.

[0123] In certain cases, half of the cells were resuspended in the thrombin component of a fibrin glue kit (Tissucol® Duo 2.0; Baxter, Madrid, Spain) prior to combination of the kit's two components, in an attemp...

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Abstract

Provided herein are novel methods and compositions utilizing adipose tissue-derived stromal stem cells for treating fistulae.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS [0001] This application is a continuation-in-part of U.S. patent application Ser. No. 11 / 065,461, filed on Feb. 25, 2005, and a continuation-in-part of U.S. patent application Ser. No. 11 / 056,241, filed on Feb. 14, 2005, the contents of which applications are herein specifically incorporated by reference in their entireties.BACKGROUND OF THE INVENTION [0002] Generally, a fistula is an abnormal connection or passageway between organs or vessels that normally do not connect. Fistulae can develop in various parts of the body. For example, types of fistulae, named for the areas of the body in which they occur, include anorectal fistula or fistula-in-ano or fecal fistula (between the rectum or other anorectal area and the skin surface), arteriovenous fistula or A-V fistula (between an artery and vein), biliary fistula (between the bile ducts to the skin surface, often caused by gallbladder surgery), cervical fistula (abnormal opening in the cervix)...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61K35/12C12N5/0775A61L27/38A61P17/02
CPCA61K35/12A61L24/106A61L27/3604A61L27/3834A61B17/1659A61L27/3895C12N5/0667C12N2533/56A61K35/35A61L27/3839A61P1/00A61P1/04A61P1/12A61P1/16A61P11/00A61P11/02A61P13/10A61P15/00A61P15/02A61P17/00A61P17/02A61P25/00A61P27/16A61P43/00A61P9/00A61P9/14C12N5/0662A61K35/36A61L27/38A61K45/06A61L24/0015A61L27/3869A61L2300/64C12N2509/00C12N2510/00A61B17/32A61B2017/320008
Inventor MIGUEL, MARIA GEMA FERNANDEZDE LA PENA, MANUEL ANGEL GONZALEZCASTRO, ROSA ANA GARCIAARRANZ, MARIANO GARCIAGARCIA-OLMO, DAMIAN
Owner AUTONOMOUS UNIVERSITY OF MADRID
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